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Updated: May 11, 2026
08:14
In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions
Published on: July 17, 2019
Cleavable trifunctional biotin reagents for protein labelling, capture and release
Yinliang Yang1, Steven H L Verhelst
1Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany.
Chemical Communications (Cambridge, England)
|May 8, 2013
Summary
New trifunctional biotin reagents with cleavable linkers improve protein enrichment. A Dde-based linker enables mild protein release and a masked trypsin site removes most of the tag during digestion.
Area of Science:
- Biochemistry
- Proteomics
- Chemical Biology
Background:
- Protein enrichment is crucial for proteomic analysis.
- Existing biotinylation reagents may lack efficient and mild cleavage mechanisms.
- The development of novel reagents is needed to improve target protein recovery and purity.
Purpose of the Study:
- To evaluate trifunctional biotin reagents with cleavable linkers for protein enrichment.
- To assess the efficiency of protein release under mild conditions.
- To investigate the utility of a masked trypsin cleavage site for tag removal.
Main Methods:
- Synthesis and application of trifunctional biotin reagents.
- Utilizing a Dde-protected linker for controlled cleavage.
- Employing a masked trypsin cleavage site within the biotin tag.
- Assessing protein release efficiency and tag removal post-digestion.
Main Results:
- The Dde-based linker facilitated efficient protein target release under mild conditions.
- The masked trypsin cleavage site effectively removed the majority of the biotin tag during tryptic digestion.
- The trifunctional biotin reagents demonstrated utility in protein enrichment strategies.
Conclusions:
- Trifunctional biotin reagents with Dde-cleavable linkers are effective for protein enrichment.
- Mild cleavage and efficient tag removal are key advantages of this approach.
- These reagents offer an improved method for preparing protein samples for analysis.
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